Neonatal screening is a proven public health service. Innovative methods such as tandem mass spectrometry have greatly increased the number of disorders effectively identified in newborn screening laboratories. To continue expanding the number of treatable disorders identified in newborns, a first tier, automated, multiplex molecular assay will be developed. The assay is based on DNA obtained from the universally collected neonatal blood card. A multiplex PCR assay will be developed to amplify DNA sequences that are diagnostic for the following disorders: sickle cell hemoglobinopathies S, C, and E alleles; alpha-1-Antitrypsin Deficiency S allele and Z allele; Factor V Leiden A1691G; and Hemochromatosis G845A. Analysis of PCR products will be performed using a low-density oligonucleotide array. The low-density oligonucleotide array will simultaneously distinguish both wild type and mutant alleles at the loci described. Data from the arrays will be processed using a BioScan Proximal Imaging System. Completing the Specific Aims of this proposal will demonstrate the feasibility of a first tier molecular screening protocol. The ability to simultaneously detect several disorders will make comprehensive newborn screening more affordable and a more effective public health service. PROPOSED COMMERCIAL APPLICATIONS: Neo Gen Screening provides a supplemental screening service, beyond that mandated by the state. Expanding the number of treatable disorders detected will make the service more comprehensive and thus more attractive to health care institutions. The screening services proposed are marketable for newborn screening and high-risk screening.